Project Summary/Abstract: Lung cancer is the most deadly cancer in the United States. Research Project 2 (P2) will accelerate advances toward the accurate diagnosis of the most common type of lung cancer, non- small cell lung cancer (NSCLC), via development of multi-scale nanotechnologies to interrogate bloodborne circulating proteins, autoantibodies, circulating tumor cells (CTCs), circulating tumor microemboli (CTM), and CTC-derived nucleic acid signatures. The proposed nanotechnology platforms strategically bring experts from diverse fields of materials science and electrical engineering (Project Leader Dr. Shan Wang), microfluidics (Co-Investigators Drs. Stephen Quake and Utkan Demirci), clinical oncology (Project Clinicians Drs. Heather Wakelee and Viswam Nair), business (Significant Contributor, Mr. Luis Carbonell, MBA), and bioinformatics and statistics (Collaborator and Consultant Drs. Olivier Gevaert and Jarrett Rosenberg) together to create rapid, accurate, and robust diagnostic tools for NSCLC patients. We endeavor to create or develop nanotechnologies ranging from magneto-nanosensors for blood-based proteomics of cancer and tumor cell enrichment with magnetic separation using magnetic sifting and magnetic levitation cell sorting, to molecular characterization using targeted multiplexed gene expression and next-generation sequencing for CTCs and CTM at the single- cell level. Our platforms focus not only on diagnosing cancer, but also on pinpointing the course and pace of cancer progression and evolution under treatment. Within the first two years, we will develop the proposed nanotechnologies ? magneto-nanosensors, magnetic separation devices, single-cell gene expression assays and gene sequencing to incorporate putative cancer biomarkers ? and then test them on two cohorts of NSCLC patients: those in early stage for diagnosis, and those in advanced stages for therapy selection and monitoring. We have access to large cohorts of clinical samples from patients provided by Stanford Hospital and MD Anderson Cancer Center. These devices will not be tested in isolation, but rather integrated with patients? clinical and imaging data from CT (computed tomography) and PET (Positron Emission Tomography)?CT, which are a crucial part of the current standard of care for diagnosis and surveillance of lung cancer. We anticipate that the development of novel nanotechnologies and pertinent methods as applied to selected bloodborne nucleic acid, proteomic, and cellular biomarkers will enable the detection of early-stage NSCLC. Unprecedented insights into targeted therapy selection and monitoring for late-stage NSCLC patients will be documented to more effectively manage patients than today?s standard of care. These nanotechnologies will enable blood biomarker analyses to be widely adopted in clinics.